Nucleic acid sequencing techniques are widely used in biological applications, for a variety of different sample types. Indeed, sequencing techniques, including those considered “next-generation” have substantial utility in forensics, diagnostics and other application. Next-generation sequencing methods are expensive, though, with a single analysis often running on the order of several thousand or several tens of thousands of dollars. It can be helpful to measure the template content of a sequencing preparation prior to providing it to a sequencing instrument for analysis, so that optimum sequencing parameters can be met. Ascertaining an amount of template in a preparation to be subject to sequencing can avoid sequencing errors, minimize waste and help improve the quality of sequencing reads that are obtained during sequencing. Many sequencing preparations utilize reagents that contain inhibitors that inhibit quantitative nucleic acid analysis suitable for quantifying the amount of template in a sequencing reaction preparation, including quantitative polymerase chain reaction (qPCR).